Name: drosophila larval nmj immunohistochemistry
Uploaded: 2009-03-28T00:00:00
Description: Watch this Scientific Journal Video about Drosophila Larval NMJ Immunohistochemistry at JoVE.com

Antes de começar a <ol><li> Prepare as seguintes soluções: PBT (0,1% Triton X100 em 1X PBS), PBTB (0,2% BSA em PBT), e PBTN (NGS 2% em PBTB). </li><li> Dissecar as larvas Drosophila. Consulte Drosophila larval Dissection MNJ. </li></ol> Imuno-histoquímica <ol><li> Mover as larvas a um tubo de 1,5 ml contendo PBT. Lavar as larvas duas vezes por 15 minutos no PBT. Nota: para lavar, coloque o tubo de 1,5 ml em um misturador nutator. Remover o líquido com um pippeto...

<ol>
	<li>Jan, L. Y., Jan, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+of+the+larval+neuromuscular+junction+in+Drosophila+Melanogaster.">Properties of the larval neuromuscular junction in Drosophila Melanogaster.</a> J. Physiol. 262, 189-214 (1976).</li><li>Johansen, J., Halpern, M. E., Johansen, K. M., Keshishian, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stereotypic+morphology+of+glutamatergic+synapses+on+identified+muscle+cells+of+Drosophila+larvae.">Stereotypic morphology of glutamatergic synapses on identified muscle cells of Drosophila larvae.</a> J Neurosci. 9, 710-725 (1989).</li><li>Kesheshian, H., Broadie, K., Chiba, A., Bate, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Drosophila+neuromuscular+junction:+a+model+system+for+studying+synaptic+development+and+function.">The Drosophila neuromuscular junction: a model system for studying synaptic development and function.</a> Annu Rev Neurosci. 19, 545-575 (1996).</li><li>Collins, C. A., DiAntonio, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+development:+insights+from+Drosophila.">Synaptic development: insights from Drosophila.</a> Curr Opin Neurobiol. 17 (1), 35-42 (2007).</li></ol>

Imuno-histoquímica (IHQ) é vital para o estudo da biologia MNJ, pois permite a visualização do MNJ. Isto é conseguido usando anticorpos que reconhecem a membrana neuronal (por exemplo, HRP), o presynapse (por exemplo, CSP, SYT), e / ou o postsynapse (por exemplo, DLG). Moléculas sinalizadoras, proteínas estruturais, ou novas proteínas de interesse também pode ser manchada. Então, os genes podem ser mutado ou missexpressed e IHC pode detectar perturbação da estrutura sináptica e / ou de sinalização neurona...

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drosophila larval nmj immunohistochemistry

O<em&gt; Drosophila</em&gt; Junção neuromuscular (MNJ) é um sistema modelo estabelecido utilizada para o estudo do desenvolvimento e plasticidade sináptica. O uso generalizado do<em&gt; Drosophila</em&gt; Motor do sistema é devido a sua alta acessibilidade. Ele pode ser analisado com uma única célula de resolução. Há 30 músculos por hemisegment cuja disposição dentro da parede do corpo periféricos são conhecidas. Um total de 31 neurônios motores atribuem a estes músculos em um padrão que tem alta fidelidade. Utilização da biologia molecular e genética, pode-se criar animais transgênicos ou mutantes. Então, pode-se estudar as conseqüências do desenvolvimento sobre a morfologia ea função do MNJ. Imuno-histoquímica pode ser usado para imagem claramente os componentes do MNJ. Neste artigo, vamos demonstrar como usar a coloração de anticorpos para visualizar a<em&gt; Drosophila</em&gt; Larval MNJ.

Watch this Scientific Journal Video about Drosophila Larval NMJ Immunohistochemistry at JoVE.com

Drosophila Larval NMJ Immunohistochemistry

Este protocolo demonstra como executar imunohistoquímica em dissecados larva Drosophila.

Drosophila larval MNJ Imunohistoquímica

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use ...

Research

JoVE Journal

Biology

Drosophila Larval NMJ Immunohistochemistry

Dissection of Larval CNS in Drosophila Melanogaster

Dissecção do CNS Larval em Drosophila melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

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Thin Sectioning of Slice Preparations for Immunohistochemistry

Seccionamento fina de Preparações Slice para Imunohistoquímica

Many investigations in neuroscience, as well as other disciplines, involve studying small, yet macroscopic pieces or sections of tissue that have been ...

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Drosophila larval Dissection MNJ

Electrophysiological Methods for Recording Synaptic Potentials from the NMJ of Drosophila Larvae

Métodos eletrofisiológicos de gravação Potenciais Synaptic do MNJ de larvas de Drosophila

In this video, we describe the electrophysiological methods for recording synaptic transmission at the neuromuscular junction (NMJ) of Drosophila larva. ...

Historical View and Physiology Demonstration at the NMJ of the Crayfish Opener Muscle

Ver histórico e demonstração Fisiologia da MNJ da Muscle Opener Crayfish

Here we present some of the key important discoveries made with the opener neuromuscular (NMJ) preparation of crustaceans and illustrate that there is ...

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

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We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

Dissection and Staining of Drosophila Larval Ovaries

Dissection and Staining of Drosophila Larval Ovaries

Dissecção e coloração de Drosophila Larval Ovários

Many organs depend on stem cells for their development during embryogenesis and for maintenance or repair during adult life. Understanding how stem cells ...

Drosophila Pupal Abdomen Immunohistochemistry

Drosophila Pupal Abdomen Immunohistochemistry

Drosophila Imunohistoquímica Abdomen Pupal

The Drosophila pupal abdomen is an established model system for the study of epithelial morphogenesis and the development of sexually dimorphic ...

Examination of Drosophila Larval Tracheal Terminal Cells by Light Microscopy

Examination of Drosophila Larval Tracheal Terminal Cells by Light Microscopy

Exame<em&gt; Drosophila</em&gt; Larval células terminais traqueais por microscopia de luz

Cell shape is critical for cell function. However, despite the importance of cell morphology, little is known about how individual cells generate specific ...

Sonication-facilitated Immunofluorescence Staining of Late-stage Embryonic and Larval Drosophila Tissues In Situ

Sonication-facilitated Immunofluorescence Staining of Late-stage Embryonic and Larval Drosophila Tissues In Situ

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Studies performed in Drosophila melanogaster embryos and larvae provide crucial insight into developmental processes such as cell fate specification and ...